Seriphidium herba-alba (Asso): A comprehensive study of essential oils, extracts, and their antimicrobial properties

Seriphidium herba-alba (Asso), a plant celebrated for its therapeutic qualities, is widely used in traditional medicinal practices throughout the Middle East and North Africa. In a detailed study of Seriphidium herba-alba (Asso), essential oils and extracts were analyzed for their chemical composition and antimicrobial properties. The essential oil, characterized using mass spectrometry and retention index methods, revealed a complex blend of 52 compounds, with santolina alcohol, α-thujone, β-thujone, and chrysanthenone as major constituents. Extraction yields varied significantly, depending on the plant part and method used; notably, methanol soaking of aerial parts yielded the most extract at 17.75%. The antimicrobial analysis showed that the extracts had selective antibacterial activity, particularly against Staphylococcus aureus, and broad-spectrum antifungal activity against organisms such as Candida albicans and Aspergillus spp. The methanol-soaked extract demonstrated the strongest antimicrobial properties, indicating its potential as a natural antimicrobial source. This study not only underscores the therapeutic potential of Seriphidium herba-alba (Asso) in pharmaceutical applications but also sets a foundation for future research focused on isolating specific bioactive compounds and in vivo testing.

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Introduction
White wormwood, known scientifically as Artemisia herba alba, is a perennial shrub indigenous to North Africa and the Middle East.It is widely recognized for its use in traditional medicine.Numerous studies have explored its diverse pharmacological properties, particularly focusing on its antibacterial and antifungal effects [1].The essential oil extracted from this plant is rich in a variety of volatile compounds, which are believed to be the key contributors to its medicinal effectiveness [2].
Among the critical bioactive components found in Artemisia herba alba are phenolic and saponin compounds, typically derived from its aerial parts.These components demonstrate notable antimicrobial activities.However, their yield and composition are subject to variation and are influenced by factors such as the specific plant parts used, and the extraction methods applied [2,3].Notably, the crude phenolic extracts from this species have shown significant effects against bacteria and fungi [4], which can be attributed to their antioxidant properties and their ability to disrupt the cell membranes of microbes [5,6].
In response to the growing problem of antibiotic-resistant bacteria, research efforts have intensified to discover new antimicrobial agents from plants [7].Artemisia herba alba extracts have been found to be effective against both gram-positive and gram-negative bacteria in various studies, highlighting their potential in combatting resistant bacterial strains, including Staphylococcus aureus and Pseudomonas aeruginosa [8].
The antifungal capabilities of Artemisia herba alba are equally significant, particularly considering the limitations and side effects of many synthetic antifungal drugs.Research has shown that Artemisia extracts can inhibit fungi such as Candida albicans, which cause infections in immunocompromised individuals [9], and their efficacy against molds such as Penicillium and Aspergillus suggests their potential use as natural food preservatives [7].
However, despite extensive studies, there is still much to learn about the comprehensive antimicrobial properties of Artemisia herba alba.Many research efforts have concentrated on isolated compounds or specific extraction techniques, often without comparing different extracts and their bioactivities [10,11].In addition, there is an ongoing need to link the chemical makeup of these extracts to their antimicrobial effectiveness for a clearer understanding of their action mechanisms.
The primary goals of this study include: analyzing the chemical composition of Artemisia herba alba's essential oils and extracts and identifying their major and minor components; comparing extraction yields from various plant parts using different methods; assessing the antibacterial efficacy of these extracts against a range of gram-positive and gram-negative bacteria; evaluating the antifungal effectiveness against common yeasts and molds; and examining the relationship between the chemical profiles of the extracts and their antimicrobial activity.This study deepen our understanding of Artemisia herba alba's pharmacological potential, emphasizing its traditional medicinal role and its possible modern therapeutic applications.

Plant collection
Artemisia herba alba specimens were collected from the Botanical Garden of Hashemite University, Zarqa, Jordan.Prof. A. El-Oqlah performed taxonomic verification at the Biological Sciences Department, Yarmouk University, Irbid, Jordan.A representative sample was archived at the Medicinal Chemistry and Pharmacognosy Department, Jordan University of Science and Technology.

Microbial strains
In this study, the bacterial strains tested encompassed both gram-positive and gram-negative strains.
Specifically, Staphylococcus aureus represented gram-positive bacteria, whereas the gram-negative group included Escherichia coli, Pseudomonas aeruginosa, Klebsiella pneumoniae, and Proteus mirabilis.In addition, the pathogenic yeast Candida albicans and various pathogenic molds such as Penicillium, Fusarium, Aspergillus, and Rhizopus species were examined.These microbial strains were obtained from the microbiology laboratory at Al-Bashir Hospital in Amman, Jordan.

Preparation of plant material
The plant was air-dried to a constant weight in a shaded area and then pulverized through a 0.25 mm sieve using a Wiley mill.The resulting powder was stored in airtight glass containers at 5°C until extraction.

Soxhlet extraction
Aerial parts and flowers (20 g) were subjected to Soxhlet extraction using petroleum ether for phenolic compounds, methanol for saponins, n-hexane for lipids and terpenoids, ethyl acetate for more polar substances, and ethanol for polar compounds.Each solvent extract was concentrated at 40-50 o C under vacuum, and the dry extracts were weighed and stored at -20°C.

Cold Extraction
Plant parts (50 g) were macerated in ethanol and methanol and separately stored for three weeks.
Homogenization was concentrated at 40-50 o C under reduced pressure.The residues were weighed and stored at -20°C.

Phenolic compound isolation
Twenty grams of powder of the aerial part was extracted with 200 mL petroleum ether for 12 h at 45 o C, then filtered with cheesecloth, and the residue was collected.The residue was then extracted with 100 mL of absolute methanol for 6 h at 45 o C. The methanolic extract was filtered and then concentrated to a small volume by evaporation under vacuum at 45 o C. Fifty milliliters of ether were added to 50 mL of 10% sodium hydroxide and extracted using a separatory funnel.The aqueous layer was removed.Concentrated HCl was added to the remaining aqueous layer for acidification.Butanol (3X20 mL) was used to extract the aqueous layer using a separatory funnel.The same aqueous layer was extracted with petroleum ether (3x20 mL).Finally, the ether extract was dried using an evaporator.The butanol extract was added to the flask and evaporated using an evaporator [12].The phenolic residue was weighed and stored in a freezer until further examination.
Detection was performed using a drop of 5% (w/v) FeCl2 solution in distilled water added to the plant extract.Green was observed, confirming the detection of phenol.Detection can also be performed using an ethanolic KOH solution, which gives a yellow to phenolic compounds [12].

Saponin fraction isolation
Twenty grams of dried powder was shaken vigorously for 15 min in a 500-mL round-bottom flask (screwcap) with chloroform (3x70 mL) to exclude fatty substances.The extract was left separate, and the supernatant was completely decanted (filtration).Then, 80% methanol (200 mL) was added to the flask and boiled for 24 h using the Soxhlet method.The procedure involved filtering the extract into a 500 mL beaker using either cheesecloth or filter paper immediately after extraction.Following this, the filtrate was subjected to vacuum evaporation at 45 o C. The remaining residue was then dissolved in 100 mL of distilled water.
Subsequently, the water-soluble portion of the residue was extracted using butanol and applied in three separate 50 mL quantities.The sterilized filtrate was concentrated under vacuum at 40-45 o C until a dry residue was obtained.The residue was weighed and stored in a freezer at -21 o C until further examination [12].
The detection of saponin compounds was performed by adding a drop of chlorosulfonic acid to the extract.A reddish-brown color was observed, confirming saponin production [12].

Water extraction
Two methods were used to obtain water extraction: The first method was according to An et al. [13].
Water extraction was performed by pouring boiling distilled water (50 mL) into 5 g of dried powder.The mixture was left to stand for 15-20 min.The mixture was then filtered through a layer of cheesecloth, and the resulting filtrate was centrifuged at 6000 rpm for 15 min.The supernatant was removed and filtered through a Millipore microfilter (0.2 µm).The sterilized filtrate was concentrated under vacuum at 40-45 o C until a dry residue was obtained.The residue was weighed and stored in a freezer at -21 o C until further examination.
The second method was performed according to Al-Charchafchi et al. [14].As mentioned previously, 50 mL of boiling distilled water was poured onto 5 g of dried powder.The process involved boiling the mixture for 10 min.After boiling, it was filtered through a layer of cheesecloth.The filtrate obtained from this step was then centrifuged at a speed of 6000 revolutions per minute for a duration of 15 min.After centrifugation, the supernatant was carefully separated and subsequently passed through a Millipore microfilter with a pore size of 0.2 µm for further purification.The sterilized filtrate was concentrated under vacuum at 40-45 o C until a dry residue was obtained.The residue was weighed and stored in a freezer at -21 o C until further examination.carrier gas helium at 1 mL/min; injection of 0.2 µL (10% hexane solution); split ratio 1:30.The identification of the components was based on comparing the retention times with those of authentic samples, comparing their linear retention indices relative to the series of n-hydrocarbons, and by computer matching against commercial (NIST 98 and ADAMS 95) and home-made library mass spectra built from pure substances and components of known essential oils [15,16].

Microbial inoculum and growth conditions
The bacterial inoculum was prepared from a nutrient broth culture incubated for 24 h at 37 o C. The suspension density was adjusted to approximately 10 4 colony-forming units per mL by comparing its turbidity with that of a McFarland 0.5 BaSO4 standard.The standard was prepared by adding 0.5 mL of 0.048 M BaCl2 to 99.5 mL of 0.36 N H2SO4.Aliquots of 4 to 6 mL were dispensed into screwcap tubes and stored in the dark at room temperature.The bacterial suspension and McFarland tube were held side by side to adjust the turbidity and viewed against a backed background.The suspension was supplemented or diluted as required.
The antimicrobial activity of various plant extracts such as n-hexane, ethyl acetate, and ethanol extracts, as well as phenols, saponin, ethanol-soaked, methanol-soaked, and water extracts extracted from various parts of Artemisia herba alba, were analyzed separately using the disk agar diffusion method [55] and evaluated against the microorganisms mentioned above.

Disk agar diffusion method
The disks were impregnated with the various plant extracts to be tested, allowed to dry, and then placed on the plates within an hour of pouring.Before disk placement, the plate surface was inoculated with a swab dipped in the standardized bacterial suspension.The surface of the plates was wiped in three directions to ensure an even and complete distribution of the inoculum throughout the plate.Discs were placed on plates and incubated at 37 o C for 24 h [17,18].
The diameter of the inhibition zones was measured in mm.For this experiment, each test was conducted in triplicate to ensure reliability and accuracy.The average of these three replicates was then calculated to obtain a more accurate and representative result.Standard antibiotics such as vancomycin 30 µL for S. aureus, streptomycin 10 µL for E. coli, K. pneumoniae, and P. mirabilis, carbenicillin 100 µL for P. aeruginosa, and amphotericin B 9.6 µg for fungi (yeast and molds) were used as appositive controls, and phosphate-buffered saline was used as a negative control.

Minimum inhibitory concentration (MIC) determination
In this study, different crude extracts of Artemisia herba alba were used to determine the minimum inhibitory concentration (MIC) against various microorganisms.To prepare each extract for testing, 100 mg of the extract was first dissolved in 1 mL of dimethyl sulfoxide (DMSO, supplied by Sigma).A series of twofold dilutions was then created.This was accomplished by adding 1 mL of the dissolved extract to test tubes containing an equal volume (1 mL) of phosphate-buffered saline (PBS) with a pH of 6.8.Bacterial cell suspensions were prepared and adjusted to give a final inoculum concentration of approximately 1 x 10 6 cells per mL (Spectronic Instruments, USA).The extract buffer tubes were inoculated with 0.5 mL of the prepared inoculum and incubated at 37 o C. The tubes were read after 24 h, and 0.1 mL of each test tube was spread on the surface of a nutrient agar plate and incubated at 37 o C for 24 h to count CFU/mL.

Chemical profile of Artemisia herba alba essential oil: main components and retention indices
Table 1 provides a detailed chemical profile of the essential oil of Artemisia herba alba and a complex series of 52 identified compounds.These were predominantly identified using mass spectrometry and retention index methods, with some verified using pure reference compounds.The oil was characterized by several main components, including santolina alcohol, α-thujone, β-thujone, chrysanthenone, cis-chrysanthenyl acetate, 1,8-cineole, camphor, and limonene, which were present in significant amounts ranging from 6.34% to 18.12%.These key components were instrumental in defining the aromatic and potential therapeutic profiles of the oil.The listed compounds exhibited various polarities and volatilities, as indicated by their linear retention indices on the apolar and polar columns.While most components were in lower concentrations, some compounds were reported without reported percentages, suggesting trace amounts or unquantified presence.
Overall, the essence of Artemisia herba alba is a rich complex of terpenes, alcohols, and ketones, reflecting the typical heterogeneity of essential oils used in various applications from the medical to the perfume industry.Extraction yields from Artemisia herba alba: comparative analysis of phenolic and saponin compounds Table 2 shows the extraction yields from different parts of the Artemisia herba alba plant, with all initial samples weighing 20 g.The extraction methods examined resulted in different extract weights, which were categorized according to the type of extract and the part of the plant used.
For crude phenolic extracts, aerial parts with flowers gave a variable weight percentage between 4.40% and 13.80%, with an average of 9.02%.The median weight percentage for these extracts was 8.10%, with the most common value being 5.15%.In contrast, immature flowers alone gave a significantly higher yield of crude phenolic extract of 13.8%, whereas mature flowers gave a yield of 8.1%.
Crude saponin extracts were also derived from different plant parts.The combined aerial parts with flowers provided a mean extract weight of 5.89%, with a range of 4.95%-7.32%and a median of 5.66%.
However, the aerial parts without flowers yielded  showing a particularly significant inhibition zone and a corresponding MIC value.K. pneumoniae was similarly affected by extracts 10 and 11, with notable inhibition zones and MIC values, although the MIC for extract 10 against this bacterium was relatively high at 50 µg/mL.
The positive control, which likely represents a standard antibiotic, consistently inhibited all tested bacterial strains, affirming the validity of the assay.Conversely, the negative control exhibited no antibacterial activity, as expected for a control.Including standard error values with the inhibition zones suggests that the experiments were conducted multiple times to determine reliability, reflecting the variability often inherent in biological testing.
Overall, the results from Table 3 demonstrate that Artemisia herba alba extracts have selective antibacterial properties, with certain extracts, particularly extract 10, showing broad-spectrum effectiveness.
The variation in activity across different extracts highlights the importance of extract type and specific bacterial strain when considering the potential therapeutic applications of these plant extracts.

Antifungal potency of Artemisia herba alba extracts in inhibition zones and minimum inhibitory concentrations
Table 4 provides a comprehensive overview of the antifungal activities of various extracts from Artemisia herba alba, as determined by the agar diffusion method and MIC tests against various fungi, including yeasts and molds.
The extracts showed varying degrees of efficacy against C. albicans, a common yeast, with extract 6 exhibiting the most potent activity, as evidenced by a 23-mm inhibition zone and an MIC value of 3.125 µg/ml.
Other extracts, such as extract 2, demonstrated significant activity with a 20-mm zone and a higher MIC of 25 µg/mL.Extracts 4, 12, and 13 inhibited C. albicans growth.
In the context of molds, specifically Penicillium spp., only a subset of the extracts, notably extract 10, showed substantial inhibitory action with a 23 mm zone and a low MIC, indicating robust antifungal properties.
Conversely, the majority of the extracts were ineffective against these mold species.
When tested against Fusarium spp., another mold type, several extracts, particularly extract 9, were effective, displaying a 21 mm inhibition zone and a MIC of 3.125 µg/mL, suggesting a strong antifungal effect at a low concentration.Similar activity patterns were observed against Aspergillus spp., with extract 9 again showing the highest activity marked by an 18 mm zone and a low MIC.
The activity against Rhizopus spp., a mold known for its rapid growth, was notable in extracts 5, 8, 9, 10, and 11, with extracts 9 and 10 displaying the largest zones of inhibition at 24 mm.The MIC values for these extracts varied, indicating differences in their effectiveness in inhibiting fungal growth.
As benchmarks, the positive control's consistent inhibition across all fungi confirmed the assay's effectiveness.In contrast, the negative control confirmed the absence of inherent antifungal properties in the medium used for testing.The standard error values associated with the inhibition zones suggest that the measurements were repeated to ensure precision, which is typical in biological assays to account for variability.
Overall, the findings from

Discussion
The present study explores the field of ethnopharmacology and phytochemistry, focusing on Artemisia herba alba, a medicinal plant.This study aligns with the growing interest in natural products for their potential therapeutic applications, particularly as antimicrobial agents.It delves into the extraction, chemical analysis, and evaluation of antimicrobial properties of various extracts from Artemisia herba alba, a plant well-known in traditional medicine.
This study also contributes to new insights into the chemical composition and bioactivity of Artemisia herba alba, particularly regarding its essential oils and extracts.The study stands out in its comprehensive approach, examining a wide range of extracts and their efficacy against both bacterial and fungal pathogens.It introduces new data on the efficacy of different extraction methods, the specific bioactivity of various extracts, and their minimum inhibitory concentrations against a spectrum of microorganisms.This includes both common pathogenic bacteria and fungi, offering a broad perspective on the plant's antimicrobial potential.
The chemical composition of the essential oil (Table 1) reveals a rich tapestry of volatile compounds, with α-thujone, β-thujone, and santolina alcohol as the predominant constituents.These findings align with those of Ouguirti et al. [19] and Houti et al. [20], who reported similar profiles in their studies of Artemisia species, emphasizing the consistency in essential oil composition across different geographical locations.
The extraction yields (Table 2) varied significantly depending on the part of the plant and the extraction method, which is in accordance with the results obtained by Brendler et al. [21] and corroborates the findings of Adam et al. [22] who noted that the extraction method profoundly affects the yield and composition of plant extracts.
The antibacterial activities (Table 3) against both gram-positive and gram-negative bacteria were noteworthy, particularly for extracts 9 (ethanol) and 10 (soaked in methanol), which showed broad-spectrum activity.These results are consistent with those of Liu et al. [23] and Bisht et al. [24], who observed substantial activity of Artemisia extracts against S. aureus and P. aeruginosa.However, our results suggest higher efficacy than the minimal inhibition concentrations reported by Ahameethunisa et al. [25].
Antifungal activities (Table 4) against a range of fungi, including C. albicans and Aspergillus spp.
Showcase some extracts with significant inhibitory properties.This is particularly interesting compared with the moderate activity reported by Mehani et al. [26], suggesting that the specific strains of Artemisia herba alba used in this study may possess unique or more potent antifungal compounds.
The potent antibacterial activity of crude phenolic extracts from immature flowers against K.
pneumoniae aligns with previous studies that have documented the antibacterial properties of phenolic compounds [27,28].The stronger antifungal efficacy against C. albicans is particularly noteworthy because it adds to the evidence supporting the use of phenolics as antifungal agents [29,30].The low MIC observed for mature flower extracts against S. aureus and C. albicans is consistent with the known bioactive profiles of mature plant parts, as reported in other studies [31][32][33].
Our results regarding crude saponin extracts are consistent with the literature that acknowledges their antimicrobial properties [34,35].The findings that extracts from both the aerial parts with and without flowers possess significant antibacterial effects against S. aureus corroborate previous research [36,37], while the efficacy against C. albicans agrees with recent studies highlighting the potential of saponins in antifungal therapy [38].
The efficacy of the ethyl acetate extract against S. aureus and its moderate effect against C. albicans add to the compound's profile as a selective antimicrobial agent [39,40].In contrast, the ethanol extracts exhibited lower activity, which may be attributed to the solvent's polarity affecting the solubility of active compounds [41].
Notably, the methanol extracts demonstrated broad-spectrum activity.This agrees with the literature suggesting that methanol may be more effective at extracting antimicrobial compounds than other solvents [42,43].However, the reduced efficacy against K. pneumonia raises questions about the specific interactions between the extract's constituents and bacterial cells [44].
The consistent MIC of 50 µg/mL for extracts soaked in ethanol reflects a lower efficacy, which may be influenced by the solvent's extraction efficiency or the intrinsic resistance of the microorganisms [45].This highlights the importance of solvent choice in the extraction process, a theme well-established in phytochemical research [46][47][48].
Our study's suggestion that the extraction method substantially impacts the antimicrobial potency of Artemisia herba alba extracts is a significant contribution to the field.Methanol soaking yields the most potent extracts, which could guide future research into optimizing extraction techniques for medicinal plants [49,50].
The variability in efficacy demonstrates the complexity of plant microbe interactions and suggests that extracts can be tailored to specific microbial targets [51,52].This tailoring could lead to the development of new antimicrobial agents that are more effective against resistant strains, a critical need given the rising concern over antibiotic resistance [53,54].
It is important to note the variability in the literature regarding the antimicrobial properties of Artemisia herba alba.While Mehani et al. [26] reported lower antimicrobial activity, our study aligns more closely with the potent bioactivities observed by El-Shatoury et al. [55].Such discrepancies could be attributed to variations in plant chemotypes, extraction methodologies, or microbial strains used in the assays, as discussed by Atef et al. [31] and Barashkova et al. [56].
The results are significant in the context of increasing antibiotic resistance and the ongoing search for novel antimicrobial compounds.By highlighting the antimicrobial properties of Artemisia herba alba extracts, the study adds valuable information to the field of natural product research and antimicrobial therapy.It underscores the importance of traditional medicinal plants as potential sources of new and effective antimicrobial agents.Additionally, the research provides a framework for future studies aiming to isolate and characterize specific bioactive compounds from Artemisia herba alba, further advancing our understanding of its therapeutic potential.The study's findings could have implications in developing new, naturally derived antimicrobial drugs, which are crucial in the face of the global challenge of antibiotic resistance.

Conclusions
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columns (30 m × 0.25 mm, 0.25 µm film thickness) and set to the following conditions: temperature program of 60 o C for 10 min, followed by an increase of 5 o C /min to 220 o C; injector and detector temperatures at 250 o C; carrier gas nitrogen (2 mL/min); detector dual FID; split ratio 1:30; and injection of standards of 0.5 µL).The chemicals were identified for both columns by comparing their retention times with those of pure, authentic samples and using their linear retention indices (LRI) relative to the series of n-hydrocarbons.The relative proportions of each component, expressed as percentages, were determined by FID peak area normalization (mean of three replicates).GC/EIMS analyses were performed using a Varian CP-3800 gas chromatograph equipped with a DB-5 capillary column(30 m × 0.25 mm; coating thickness, 0.25 µm) and a Varian Saturn 2000 ion trap mass detector.Analysis conditions included: injector and transfer line temperatures at 220 and 240 o C respectively; the oven temperature programed from 60 o C to 240 o C at 3 o C /min; particularly within its essential oil.It has shown variable yet significant antibacterial and antifungal activities across different extracts.The results suggest that both the chemical composition and extraction technique are crucial in determining the efficacy of the extracts against various microorganisms.Although this study provides valuable insights into the potential use of Artemisia herba alba as a natural source of antimicrobial agents, the findings are constrained by limitations such as the lack of in vivo testing, potential seasonal and geographical variation in plant chemistry, and the need for further toxicological assessment.Future investigations are warranted to explore the full therapeutic potential and safety of these extracts and the mechanisms underlying their bioactivity.

Table 1 . Chemical composition of Artemisia herba alba essential oil.
a Linear retention index (apolar column) b Linear retention index (polar column) c trace amounts < 0.1 e identification: MS=mass spectrometry, RI=retention index, ST=pure reference compound

Table 2 . Various extracts of Artemisia herba alba. The weight of the plant parts was 20 g.
Notably, some extracts have detailed statistical data, such as crude saponin from aerial parts with flowers, indicating multiple measurements and variations in the extraction process.However, for several extracts, only single values were provided without additional statistical data, suggesting that either a single measurement was taken, or the variability needed to be documented.The efficiency and yield of the extraction process depend on both the plant part being processed and the extraction method used.The table provides an overview of how different techniques and plant parts contribute to the yield of Artemisia herba alba extracts.
5.14%, immature flowers yielded 7.32%, and mature flowers yielded the lowest yield at 4.95%.Other extraction methods were used on aerial parts with flowers, resulting in consistent yields across single measurements.The ethyl acetate and ethanol extracts had uniform weights of 9.15% and 7.67%, respectively.Soaking the aerial parts in methanol gave the highest extract weight at 17.75%, whereas ethanol soaking gave 15.53%.The two different water extraction methods each yielded 1%.

Table 3
evaluates the antibacterial efficacy of various extracts from Artemisia herba alba using agar diffusion and minimum inhibitory concentration (MIC) methods against a spectrum of bacteria.The findings reveal that the plant extracts exhibit a range of activities against both gram-positive and gram-negative bacteria, with varying levels of effectiveness.Several extracts demonstrated antibacterial action against the gram-positive bacterium S. aureus, indicated by inhibition zones measuring between 11 and 20 mm.In particular, extracts 2, 4, 5, 6, 7, 8, and 9 showed inhibition, with the most potent being extract 10.The MIC values complemented these findings, with some extracts displaying substantial bacteriostatic properties at concentrations as low as 3.125 µg/mL.Notably, extracts 1, 3, 12, and 13 did not inhibit S. aureus, suggesting that they lack the necessary components or concentrations to affect this bacterium.Regarding gram-negative bacteria, the extracts generally exhibited less activity.E. coli was resistant to most extracts, except for extract 10, which inhibited growth with an inhibition zone of 13 mm and a MIC of 3.125 µg/mL.P. aeruginosa was only susceptible to extract 1, which displayed a moderate inhibition zone and MIC indicative of moderate effectiveness.For P. mirabilis, extracts 10 and 11 were effective, with extract 10

Table 3 . Antibacterial activity of various extracts of Artemisia herba alba by agar diffusion and MIC methods.
DDM, disc diffusion method; MIC, minimal inhibition concentration; *Inhibition zone in mm ± SE (S.E., Standard error).

Table 4 . Antifungal activity of various extracts of Artemisia herba alba by agar diffusion and MIC
Table 4 underscore the potential of Artemisia herba alba extracts as antifungal agents, with certain extracts, particularly numbers 9 and 10, demonstrating broad-spectrum antifungal properties against various fungal pathogens.The data indicate that the efficacy of the extracts can be speciesspecific and suggest a need for further research to isolate and understand the active compounds contributing to these antifungal effects.